TY - CHAP
T1 - Qflow cytometry-based receptoromic screening
T2 - A high-throughput quantification approach informing biomarker selection and nanosensor development
AU - Chen, Si
AU - Weddell, Jared
AU - Gupta, Pavan
AU - Conard, Grace
AU - Parkin, James
AU - Imoukhuede, Princess I.
N1 - Publisher Copyright:
© Springer Science+Business Media LLC 2017.
PY - 2017
Y1 - 2017
N2 - Nanosensor-based detection of biomarkers can improve medical diagnosis; however, a critical factor in nanosensor development is deciding which biomarker to target, as most diseases present several biomarkers. Biomarker-targeting decisions can be informed via an understanding of biomarker expression. Currently, immunohistochemistry (IHC) is the accepted standard for profiling biomarker expression. While IHC provides a relative mapping of biomarker expression, it does not provide cell-by-cell readouts of biomarker expression or absolute biomarker quantification. Flow cytometry overcomes both these IHC challenges by offering biomarker expression on a cell-by-cell basis, and when combined with calibration standards, providing quantitation of biomarker concentrations: this is known as qFlow cytometry. Here, we outline the key components for applying qFlow cytometry to detect biomarkers within the angiogenic vascular endothelial growth factor receptor family. The key aspects of the qFlow cytometry methodology include: antibody specificity testing, immunofluorescent cell labeling, saturation analysis, fluorescent microsphere calibration, and quantitative analysis of both ensemble and cell-by-cell data. Together, these methods enable high-throughput quantification of biomarker expression.
AB - Nanosensor-based detection of biomarkers can improve medical diagnosis; however, a critical factor in nanosensor development is deciding which biomarker to target, as most diseases present several biomarkers. Biomarker-targeting decisions can be informed via an understanding of biomarker expression. Currently, immunohistochemistry (IHC) is the accepted standard for profiling biomarker expression. While IHC provides a relative mapping of biomarker expression, it does not provide cell-by-cell readouts of biomarker expression or absolute biomarker quantification. Flow cytometry overcomes both these IHC challenges by offering biomarker expression on a cell-by-cell basis, and when combined with calibration standards, providing quantitation of biomarker concentrations: this is known as qFlow cytometry. Here, we outline the key components for applying qFlow cytometry to detect biomarkers within the angiogenic vascular endothelial growth factor receptor family. The key aspects of the qFlow cytometry methodology include: antibody specificity testing, immunofluorescent cell labeling, saturation analysis, fluorescent microsphere calibration, and quantitative analysis of both ensemble and cell-by-cell data. Together, these methods enable high-throughput quantification of biomarker expression.
KW - Angiogenesis
KW - Background subtraction
KW - Heterogeneity
KW - Immuno-labeling
KW - Mixture modeling
KW - Platelet-Derived Growth Factor (PDGF)
KW - QFlow cytometry
KW - Quantitative flow cytometry
KW - Systems biology
KW - Vascular Endothelial Growth Factor (VEGF)
UR - http://www.scopus.com/inward/record.url?scp=85014029727&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85014029727&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-6840-4_8
DO - 10.1007/978-1-4939-6840-4_8
M3 - Chapter
C2 - 28238133
AN - SCOPUS:85014029727
T3 - Methods in Molecular Biology
SP - 117
EP - 138
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -